Analysis of endogenous hormone changes and gene expression related to walnut apomixis kernels formation

doi:10.3969/j.issn.1004-1524.20231390

Acta Agriculturae Zhejiangensis ›› 2025, Vol. 37 ›› Issue (2): 381-393.DOI: 10.3969/j.issn.1004-1524.20231390

• Horticultural Science • Previous Articles Next Articles

Analysis of endogenous hormone changes and gene expression related to walnut apomixis kernels formation

CHEN Feng(), CHEN Hong(), CHEN Bingquan, BAO Chunjie, ZHOU Haoliang, ZHAO Xin, GUO Laizhen

College of Forestry and Landscape Architecture, Xinjiang Agricultural University, Urumqi 830052, China

Received:2023-12-12 Online:2025-02-25 Published:2025-03-20
Contact: CHEN Hong

Abstract

Abstract:

In order to understand the molecular regulation of apomictic nucleolus development in walnuts, the apomictic kernels and fertilized kernels of ‘Xinxin 2’ walnut were used as the experimental materials in this study. The transcriptome and hormone contents such as indole-3-acetic acid (IAA), cytokinin (ZT), gibberellin (GA), brassinosteroid (BR), abscisic acid (ABA), and ethylene (ETH) were analyzed. The results showed that at 60 days after flowering, the contents of IAA, ZT, and BR in apomictic kernels were significantly (P<0.05) lower than those in normally fertilized kernels, decreasing by 28.57%, 19.59%, and 12.07%, respectively; the contents of GA, ABA, and ETH in apomictic kernels were significantly (P<0.05) higher than those in normally fertilized kernels, increasing by 34.91%, 31.50%, and 19.54%, respectively. At 80 days after flowering, the contents of IAA, ZT, and ETH in apomictic kernels were significantly (P<0.05) higher than those in normally fertilized kernels, increasing by 6.58%, 17.80%, and 7.44%, respectively; meanwhile, the contents of GA, BR, and ABA in apomictic kernels were significantly (P<0.05) lower than those in normally fertilized kernels, decreasing by 29.37%, 14.99%, and 10.65%, respectively. At 100 days after flowering, the contents of IAA, ZT, GA, BR, and ETH in apomictic kernels were significantly (P<0.05) higher than those in normally fertilized kernels, increasing by 18.87%, 30.12%, 16.96%, 3.70%, and 20.57%, respectively; the content of ABA in apomictic kernels was significantly (P<0.05) lower than those in normally fertilized kernels, decreasing by 44.85%. Transcriptome sequencing result showed that 62 differentially expressed genes (DEGs) involved in plant hormone signal transduction pathways were identified. Among them, auxin influx carrier gene (AUX1 LAX_2), B-class cytokinin response factor gene (ARR-B1), abscisic acid receptor gene (PYL2), BAK family gene (BAK1), and phosphotransferase gene (AHP4) were upregulated in apomictic kernels, whereas F-box protein gene (GID2_SLY1_2) and E3 ubiquitin ligase genes (EBF1/2_1, EBF1/2_2) were downregulated. Eight genes that related to differentially metabolites were verified by qRT-PCR and the results were consistent with the finding in transcriptome analysis. It was concluded that during the oil conversion period of walnuts, high contents of IAA, ZT, BR, ETH could promote the development of apomictic kernels. Genes such as AUX1 LAX_2, ARR-B1, PYL2, BAK1, AHP4, GID2_SLY1_2, and EBF1/2 which connect with plant hormone signal transduction pathways were participating in regulating the development of apomictic kernel development.

Key words: walnut, apomixis, kernel, endogenous hormone, transcriptome

CLC Number:

S664.1

CHEN Feng, CHEN Hong, CHEN Bingquan, BAO Chunjie, ZHOU Haoliang, ZHAO Xin, GUO Laizhen. Analysis of endogenous hormone changes and gene expression related to walnut apomixis kernels formation[J]. Acta Agriculturae Zhejiangensis, 2025, 37(2): 381-393.

Figures/Tables 10

Table 1 Primer sequences of candidate genes used for real-time fluorescence quantitative PCR

基因ID Gene ID	基因名称 Gene name	正向引物序列 Forward primer sequence (5'→3')	反向引物序列 Reverse primer sequence (5'→3')
c61859_g2	LUX1 LAX_2	CGTTACGTCGGGAGGATGAG	ATTGAGCGACGAAAAGGGGT
c60146_g1	GID2_SLY1_2	TGGCTATTGCAAAATCAGGTC	ACGCTCGAGTGTAGTAGCAAG
c66727_g2	ARR-B2	CTCACCCTTGTAGAGCGAGG	TGCAAACTCAGTGCTAAAATCAA
c65820_g2	AHP4	ACCCATCTTTCACGTGTGTG	TGCCTAATTAGCAATTTCCA
c63550_g3	EBF1/2_1	CGTTACGTCGGGAGGATGAG	CGTTACGTCGGGAGGATGAG
c64965_g1	EBF1/2_2	ACCTTCCCTGCTCCTCTCTT	GAGCCTTGTGGAAGCAAACG
c56774_g2	PYL2	TGTAATTGGGGAATGTTGCA	TGGGAGACACAATGATCGAA
c66515_g2	BAK1_2	AACTGTTGCCGGAGCTTTCT	TGGGATAACACCACATGCAGT

Fig.1 The endogenous hormone content of the apomictic kernels and fertilized kernels of ‘Xinxin 2’ walnut at different developmental stages Different letters indicate significant difference at P<0.05.

Table 2 Statistical table for quality assessment of sequencing data

样品编号 Sample ID	原始数据 Raw read	过滤后数据 Clean read	高质量碱基 Clean base/Gb	碱基质量值Quality score		整体测序错误率 Error rate/%	GC含量 GC content/%
样品编号 Sample ID	原始数据 Raw read	过滤后数据 Clean read	高质量碱基 Clean base/Gb	Q20/%	Q30/%	整体测序错误率 Error rate/%	GC含量 GC content/%
G1	52981375	50634989	7.60	95.63	90.14	0.02	45.53
G2	51860163	49469256	7.42	95.38	89.66	0.02	48.78
G3	53160610	50416068	7.56	95.85	90.36	0.02	48.96
W1	45992618	43736529	6.56	94.85	88.63	0.02	44.79
W2	48272843	44792770	6.72	94.87	88.63	0.02	46.45
W3	47683476	44944840	6.74	95.23	89.46	0.02	45.16

Fig.2 Analysis of genes related to IAA metabolism and signal transduction in the apomictic kernels and fertilized kernels of ‘Xinxin 2’ walnut G1-G3 and W1-W3 represent the three periods of fertilized kemels and apomictic kemels of ‘Xinxia 2’ walnut.The same as below.

Fig.3 Analysis of genes related to cytokinin metabolism and signal transduction in apomictic kernels and fertilized kernels of ‘Xinxin 2’ walnut

Fig.4 Analysis of genes related to gibberellin metabolism and signal transduction in apomictic kernels and fertilized kernels of ‘Xinxin 2’ walnut

Fig.5 Analysis of genes related to ethylene metabolism and signal transduction in apomictic kernels and fertilized kernels of ‘Xinxin 2’ walnut

Fig.6 Analysis of genes related to abscisic acid metabolism and signal transduction in apomictic kernels and fertilized kernels of ‘Xinxin 2’ walnut

Fig.7 Analysis of genes related to brassinosteroid metabolism and signal transduction in apomictic kernels and fertilized kernels of ‘Xinxin 2’ walnut

Fig.8 Relative expression of differentially expressed genes

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Analysis of endogenous hormone changes and gene expression related to walnut apomixis kernels formation

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